LOCUS       sequence-417144-       12156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter gene expression in B. dendrobatidis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12156)
  AUTHORS   Kalinka E, Brody SM, Swafford AJM, Medina EM, Fritz-Laylin LK
  TITLE     Genetic transformation of the frog-killing chytrid fungus 
            Batrachochytrium dendrobatidis.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Jan 23;121(4):e2317928121. doi: 
            10.1073/pnas.2317928121. Epub 2024 Jan 18.
  PUBMED    38236738
REFERENCE   2  (bases 1 to 12156)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2024-01-23"; volume: "121(4):e2317928121. 
            doi"; pages: " 10.1073/pnas.2317928121. Epub 2024 Jan 18"
COMMENT     Sequence Label: pEK24
FEATURES             Location/Qualifiers
     source          1..12156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     206..228
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     220..237
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     221..237
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1259..2284
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1259..2284
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             2300..2815
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     terminator      2837..3024
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2892..2916)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3039..3055)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3039..3055)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3052..3074)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3063..3079
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3087..3093,3094..3111,3112..3117))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3132..3153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    3330..3354
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(3636..3655)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3647..4462
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3647..4462
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3720..3739)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4557..5145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5046..5065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5299..5316
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5331..5471
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      5815..6009
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6075..7148)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(7577..8206)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     rep_origin      9367..10709
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(10330..10377)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     10750..10768
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10971..11186
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             11187..11990
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc tcattcgcca ttcaggctgc gcaactgttg ggaagggcga
      121 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      181 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
      241 aagcttgaca gtatcataaa tggttgctgt atctattgaa ttgagaatgc caataaagtt
      301 aatttgtagc aggaaaatag agcaaatatt taatcattat ttattctgaa gaggcagcca
      361 ccaagatata ttcaaatttc attctacatg aatacagctg taaatatggt gttatcacag
      421 ggcttagtga accaatggtt gatcatggaa gcaacgtcaa gactaagctc tagcactgaa
      481 atgtggaaag tgcgggtcat ctcacccata gactcgcttg tcttgtctaa aacaagattt
      541 gtcttgatca agatatatgt aacactgagt tgccatatag attgtgttta tatgaaaagt
      601 agttttccaa caagtgtagc ttggagttgg aacgagtatt tagttggaca tgagattgac
      661 gtattagcca tccatgcaag caaatccaag ccaagttgaa atgtccagag gtttgattcc
      721 ttctagagcg gagctgtggc gcattctaaa catgaaagcg tggcttggtt gctggagtcg
      781 ctagattcgc taatgttgct gaattagatt tttgcacctg ttgatttata ttagtcggat
      841 gtattgctta tcaatatttt acaattacaa ttaaacatgt agcggataga ctagcttatt
      901 gagtcttatg gtttagatac gatatacatc ccaagatatc ttgagagaag cctatgctag
      961 cacttatgga acagaaggcc agtttgccat tgaactagtt agtataaaat gggttgacag
     1021 cacatgagga ctcttacaat aaaatacgag taacttgggg tggagtgtcg atcggctagt
     1081 taaagcaatg cactgtttgt tccctggctc aatattcact ccacactttc agctagatcc
     1141 gccaccagtc atccacacga aaatttccta caaattccgg ttacttttcc ctatttcttc
     1201 tctttccatc acaagaatct atattcaagt taccccttct tactatctaa tttaattaat
     1261 gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     1321 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     1381 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     1441 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
     1501 tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     1561 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga
     1621 tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     1681 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     1741 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     1801 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     1861 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     1921 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     1981 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     2041 gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     2101 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     2161 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
     2221 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     2281 ggaaggagga ggaggatcca tggtcttcac actcgaagat ttcgttgggg actggcgaca
     2341 gacagccggc tacaacctgg accaagtcct tgaacaggga ggtgtgtcca gtttgtttca
     2401 gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc ctgagcggtg aaaatgggct
     2461 gaagatcgac atccatgtca tcatcccgta tgaaggtctg agcggcgacc aaatgggcca
     2521 gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat catcacttta aggtgatcct
     2581 gcactatggc acactggtaa tcgacggggt tacgccgaac atgatcgact atttcggacg
     2641 gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc actgtaacag ggaccctgtg
     2701 gaacggcaac aaaattatcg acgagcgcct gatcaacccc gacggctccc tgctgttccg
     2761 agtaaccatc aacggagtga ccggctggcg gctgtgcgaa cgcattctgg cgtaaaggcg
     2821 cgccacttct aaataagcga atttcttatg atttatgatt tttattatta aataagttat
     2881 aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt
     2941 attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc
     3001 tcttattgac cacacctcta ccgggaattc gtaatcatgt catagctgtt tcctgtgtga
     3061 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3121 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3241 ggtttgcgta ttggctagag caattcggcg ttaattcagt acattaaaaa cgtccgcaat
     3301 gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc
     3361 agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca
     3421 tcagtccggg acggcgtcag cgggagagcc gttgtaaggc ggcagacttt gctcatgtta
     3481 ccgatgctat tcggaagaac ggcaactaag ctgccgggtt tgaaacacgg atgatctcgc
     3541 ggagggtagc atgttgattg taacgatgac agagcgttgc tgcctgtgat ccagatcatg
     3601 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     3661 acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg atttatatgg
     3721 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg
     3781 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3841 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3901 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac
     3961 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     4021 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     4081 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     4141 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     4201 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     4261 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     4321 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     4381 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     4441 gatgctcgat gagtttttct aagaattaat tcatgatcct gcatgaccaa aatcccttaa
     4501 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4561 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4621 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4681 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4741 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4801 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4861 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4921 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4981 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5041 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5101 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5161 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5221 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5281 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     5341 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     5401 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     5461 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5521 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5581 tttcaccgtc atcaccgaaa cgcgcgaggc agggtgcctt gatgtgggcg ccggcggtcg
     5641 agtggcgacg gcgcggcttg tccgcgccct ggtagattgc ctggccgtag gccagccatt
     5701 tttgagcggc cagcggccgc gataggccga cgcgaagcgg cggggcgtag ggagcgcagc
     5761 gaccgaaggg taggcgcttt ttgcagctct tcggctgtgc gctggccaga cagttatgca
     5821 caggccaggc gggttttaag agttttaata agttttaaag agttttaggc ggaaaaatcg
     5881 ccttttttct cttttatatc agtcacttac atgtgtgacc ggttcccaat gtacggcttt
     5941 gggttcccaa tgtacgggtt ccggttccca atgtacggct ttgggttccc aatgtacgtg
     6001 ctatccacag gaaagagacc ttttcgacct ttttcccctg ctagggcaat ttgccctagc
     6061 atctgctccg tacattagga accggcggat gcttcgccct cgatcaggtt gcggtagcgc
     6121 atgactagga tcgggccagc ctgccccgcc tcctccttca aatcgtactc cggcaggtca
     6181 tttgacccga tcagcttgcg cacggtgaaa cagaacttct tgaactctcc ggcgctgcca
     6241 ctgcgttcgt agatcgtctt gaacaaccat ctggcttctg ccttgcctgc ggcgcggcgt
     6301 gccaggcggt agagaaaacg gccgatgccg ggatcgatca aaaagtaatc ggggtgaacc
     6361 gtcagcacgt ccgggttctt gccttctgtg atctcgcggt acatccaatc agctagctcg
     6421 atctcgatgt actccggccg cccggtttcg ctctttacga tcttgtagcg gctaatcaag
     6481 gcttcaccct cggataccgt caccaggcgg ccgttcttgg ccttcttcgt acgctgcatg
     6541 gcaacgtgcg tggtgtttaa ccgaatgcag gtttctacca ggtcgtcttt ctgctttccg
     6601 ccatcggctc gccggcagaa cttgagtacg tccgcaacgt gtggacggaa cacgcggccg
     6661 ggcttgtctc ccttcccttc ccggtatcgg ttcatggatt cggttagatg ggaaaccgcc
     6721 atcagtacca ggtcgtaatc ccacacactg gccatgccgg ccggccctgc ggaaacctct
     6781 acgtgcccgt ctggaagctc gtagcggatc acctcgccag ctcgtcggtc acgcttcgac
     6841 agacggaaaa cggccacgtc catgatgctg cgactatcgc gggtgcccac gtcatagagc
     6901 atcggaacga aaaaatctgg ttgctcgtcg cccttgggcg gcttcctaat cgacggcgca
     6961 ccggctgccg gcggttgccg ggattctttg cggattcgat cagcggccgc ttgccacgat
     7021 tcaccggggc gtgcttctgc ctcgatgcgt tgccgctggg cggcctgcgc ggccttcaac
     7081 ttctccacca ggtcatcacc cagcgccgcg ccgatttgta ccgggccgga tggtttgcga
     7141 ccgctcacgc cgattcctcg ggcttggggg ttccagtgcc attgcagggc cggcagacaa
     7201 cccagccgct tacgcctggc caaccgcccg ttcctccaca catggggcat tccacggcgt
     7261 cggtgcctgg ttgttcttga ttttccatgc cgcctccttt agccgctaaa attcatctac
     7321 tcatttattc atttgctcat ttactctggt agctgcgcga tgtattcaga tagcagctcg
     7381 gtaatggtct tgccttggcg taccgcgtac atcttcagct tggtgtgatc ctccgccggc
     7441 aactgaaagt tgacccgctt catggctggc gtgtctgcca ggctggccaa cgttgcagcc
     7501 ttgctgctgc gtgcgctcgg acggccggca cttagcgtgt ttgtgctttt gctcattttc
     7561 tctttacctc attaactcaa atgagttttg atttaatttc agcggccagc gcctggacct
     7621 cgcgggcagc gtcgccctcg ggttctgatt caagaacggt tgtgccggcg gcggcagtgc
     7681 ctgggtagct cacgcgctgc gtgatacggg actcaagaat gggcagctcg tacccggcca
     7741 gcgcctcggc aacctcaccg ccgatgcgcg tgcctttgat cgcccgcgac acgacaaagg
     7801 ccgcttgtag ccttccatcc gtgacctcaa tgcgctgctt aaccagctcc accaggtcgg
     7861 cggtggccca tatgtcgtaa gggcttggct gcaccggaat cagcacgaag tcggctgcct
     7921 tgatcgcgga cacagccaag tccgccgcct ggggcgctcc gtcgatcact acgaagtcgc
     7981 gccggccgat ggccttcacg tcgcggtcaa tcgtcgggcg gtcgatgccg acaacggtta
     8041 gcggttgatc ttcccgcacg gccgcccaat cgcgggcact gccctgggga tcggaatcga
     8101 ctaacagaac atcggccccg gcgagttgca gggcgcgggc tagatgggtt gcgatggtcg
     8161 tcttgcctga cccgcctttc tggttaagta cagcgataac cttcatgcgt tccccttgcg
     8221 tatttgttta tttactcatc gcatcatata cgcagcgacc gcatgacgca agctgtttta
     8281 ctcaaataca catcaccttt ttagacggcg gcgctcggtt tcttcagcgg ccaagctggc
     8341 cggccaggcc gccagcttgg catcagacaa accggccagg atttcatgca gccgcacggt
     8401 tgagacgtgc gcgggcggct cgaacacgta cccggccgcg atcatctccg cctcgatctc
     8461 ttcggtaatg aaaaacggtt cgtcctggcc gtcctggtgc ggtttcatgc ttgttcctct
     8521 tggcgttcat tctcggcggc cgccagggcg tcggcctcgg tcaatgcgtc ctcacggaag
     8581 gcaccgcgcc gcctggcctc ggtgggcgtc acttcctcgc tgcgctcaag tgcgcggtac
     8641 agggtcgagc gatgcacgcc aagcagtgca gccgcctctt tcacggtgcg gccttcctgg
     8701 tcgatcagct cgcgggcgtg cgcgatctgt gccggggtga gggtagggcg ggggccaaac
     8761 ttcacgcctc gggccttggc ggcctcgcgc ccgctccggg tgcggtcgat gattagggaa
     8821 cgctcgaact cggcaatgcc ggcgaacacg gtcaacacca tgcggccggc cggcgtggtg
     8881 gtgtcggccc acggctctgc caggctacgc aggcccgcgc cggcctcctg gatgcgctcg
     8941 gcaatgtcca gtaggtcgcg ggtgctgcgg gccaggcggt ctagcctggt cactgtcaca
     9001 acgtcgccag ggcgtaggtg gtcaagcatc ctggccagct ccgggcggtc gcgcctggtg
     9061 ccggtgatct tctcggaaaa cagcttggtg cagccggccg cgtgcagttc ggcccgttgg
     9121 ttggtcaagt cctggtcgtc ggtgctgacg cgggcatagc ccagcaggcc agcggcggcg
     9181 ctcttgttca tggcgtaatg tctccggttc tagtcgcaag tattctactt tatgcgacta
     9241 aaacacgcga caagaaaacg ccaggaaaag ggcagggcgg cagcctgtcg cgtaacttag
     9301 gacttgtgcg acatgtcgtt ttcagaagac ggctgcactg aacgtcagaa ggaaaagtgc
     9361 cacctgaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
     9421 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
     9481 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
     9541 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     9601 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
     9661 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
     9721 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
     9781 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
     9841 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
     9901 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
     9961 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
    10021 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
    10081 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
    10141 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
    10201 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt
    10261 acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa
    10321 aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag gaacttcaaa
    10381 gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat acagctcact
    10441 gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga gaagaacggc
    10501 atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta ggatgaaagg
    10561 tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat gcttccttca
    10621 gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt ctcatccttc
    10681 aatgctatca tttcctttga tattggatca tactaagaaa ccattattat catgacatta
    10741 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
    10801 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
    10861 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
    10921 aactatgcgg catcagagca gattgtactg agagtgcacc ataccacctt ttcaattcat
    10981 catttttttt ttattctttt ttttgatttc ggtttccttg aaattttttt gattcggtaa
    11041 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
    11101 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
    11161 aaacctgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
    11221 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
    11281 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
    11341 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
    11401 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca
    11461 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
    11521 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
    11581 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgctagcag
    11641 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
    11701 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
    11761 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg
    11821 cattgggtca acagtataga accgtggatg atgtggtctc tacaggatct gacattatta
    11881 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
    11941 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
    12001 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
    12061 tattacctgg catgcacata caaatggacg aacggataaa ccttttcacg cccttttaaa
    12121 tatccgatta ttctaataaa cgctcttttc tcttag
//
